Documente Academic
Documente Profesional
Documente Cultură
Abstract | CD4+ Tcells differentiate and acquire distinct functions to combat specific
pathogens but can also adapt their functions in response to changing circumstances. Although
this phenotypic plasticity can be potentially deleterious, driving immune pathology, it also
provides important benefits that have led to its evolutionary preservation. Here, we review
CD4+ Tcell plasticity by examining the molecular mechanisms that regulate it from the
extracellular cues that initiate and drive cells towards varying phenotypes, to the cytosolic
signalling cascades that decipher these cues and transmit them into the cell and to the
nucleus, where these signals imprint specific gene expression programmes. By understanding
how this functional flexibility is achieved, we may open doors to new therapeutic approaches
that harness this property of Tcells.
REVIEWS
Sense
TH1 cell
IL-2, IL-12
and IFN
Strong
TCR signal
TH2 cell
IL-2
and IL-4
TH9 cell
IL-2, IL-4
Weak
and TGF
TCR signal
TFH cell
IL-6
and
IL-21
Strong
TCR
signal ICOS
TCR
TH17 cell
IL-6, IL-21,
IL-23 and
TGF
TCR
TReg cell
IL-2
tTReg: strong signal
and
pTReg: weak signal
TGF
CD28
Programme
TCR
STAT1 T-bet
STAT4
Function
CXCR3
and CCR5
IFN
GATA3
STAT6
GFI1, IRF4 and MAF
CCR4
and CCR8
IRF4, SMAD2
and SMAD3
IL-4, IL-5
and IL-13
BCL-6
STAT3
PU.1
STAT6
CCR3,
CXCR3
and CCR6
IL-9
CXCR5
IL-21
CD40L
Activate B cells
Increase antibody
anity (SHM)
Defence against
extracellular parasites Defence against
extracellular
Mucosal autoimmunity
pathogens
Cancer regulation
ROR t
STAT3
AHR, HIF1
and ROR
CCR6
IL-17 and
IL-22
Activate
neutrophils
Activate epithelium
Defence against
extracellular
pathogens
(e.g. fungi)
FOXP3
STAT5
BACH2, eos, FOXO1
or FOXO3, NR4A,
SMAD2 or SMAD3
PD1 CTLA4
IL-10 and
TGF
Figure 1 | Polarized CD4+ Tcell subsets. Each CD4+ Tcell subset can be defined by their distinct abilities to sense
(red), programme (orange) and function (blue) in the control of specific pathogens or immuneNature
pathologies.
The
inductive
Reviews
| Immunology
cytokines, polarizing transcription factors and cytokines or chemokine receptors that are characteristic of each subset are
shown, along with their association with specific forms of immune defence. AHR, aryl hydrocarbon receptor; BATF,
Bcell-activating transcription factor; BCL6, B cell lymphoma 6; CCR, CC-chemokine receptor; CD40L, CD40 ligand;
CTLA4, cytotoxic T lymphocyte antigen 4; CXCR, CXC-chemokine receptor; EOMES, eomesodermin; FOXO, forkhead
boxO; FOXP3, forkhead box P3; GATA3, GATA-binding protein 3; GFI1, growth-factor independent 1; HIF1,
hypoxia-inducible factor 1; ICOS, inducible T cell co-stimulator; IFN, interferon-; IL, interleukin; IRF4, interferonregulatory factor 4; MAF, macrophage-activating factor; NR4A, nuclear receptor 4A; PD1, programmed cell death1;
pTRegcell, peripherally derived regulatory T cell; RAR, retinoic acid receptor; ROR, retinoic acid receptor-related orphan
receptor; RUNX3, runt-related transcription factor 3; SHM, somatic hypermutation; STAT, signal transducer and activator
of transcription; TCF1, T cell factor 1; TCR, T cell receptor; TFH, T follicular helper; TGF, transforming growth factor;
TH,Thelper; TReg cell, regulatory T cell; tTReg cell, thymus-derived regulatory T cell.
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
Box 1 | Lineage stability or plasticity?
The T helper 1 (TH1) versus TH2 lineage hypothesis was ground-breaking because it
established a paradigm for Tcells to differentiate and acquire specific functions.
However, it also diverted attention from the functional plasticity of Tcells. Even at that
time, demonstrations of the coexpression of interferon (IFN) and interleukin4 (IL4)
from human Tcell clones was prevalent199,200, and single cell analysis showed
coexpression of TH1- and TH2type cytokines, especially early after the initial stimulation
of naive Tcells201. Nevertheless, the capacity of CD4+ T cells to exhibit stably polarized
phenotypes both invitro (under specific polarizing conditions) and invivo (inindividuals
with immunopathologies) supported the hypothesis that distinct Tcell lineages can
develop that supersede plasticity2,3,201. However, as discussed in this Review, it is now
apparent that Tcells can exhibit both polarized Tcell functions and phenotypic plasticity
throughout their lifespan. Thus, terminology that encompasses both of these traits is
important6,10,116. We favour the original descriptor polarized Tcell subset because it
implies an active process with degrees of divergence and leaves room for flexibility in
phenotypes over time. This sets the stage for Tcells transitioning between phenotypes,
not as a consequence of failed maintenance of aspecified programme, as implied by
loss of lineage stability, but owing to a programmed retention of plasticity or the
capacity of Tcells to dynamically control their function in response to changing
contexts. Thus, Tcell differentiation closely resembles the plasticity observed in the
nervous system, wherein programmes are not hardwired but are flexible in response to
changing environmental cues. The use of terms such as lineage, specification and master
regulator are terms best aligned with developmental programmes, such as in
Caenorhabditiselegans, in which the fate of each cell is predetermined and irreversible.
REVIEWS
TH2 or TFH cell differentiation can dominate5558. By con
trast, weak TCR signalling, by transient or low-affinity
TCR interactions, favours the induction of FOXP3
expression59,60. Importantly, TCR signal strength drives
the first divergence of inflammatory versus regulatory
subsets during Tcell development in the thymus. An
increased strength of signal, which is imparted by TCRs
that recognize self-antigen, imprints a distinct epigenetic
state and induces FOXP3 expression to generate the
TRegcelllineage61,62.
TCR and costimulatory signals have key roles in
the development of polarized Tcell subsets, but less is
known about the role of TCR stimulation in driving plas
ticity between subsets. Weaker TCR stimulation during
priming invivo allows for greater plasticity in second
ary responses63, but varying the strength of signal dur
ing recall responses also redirects T helper cell subsets,
with stronger signals promoting TH2 cell phenotypes
in memory TH1 cells64. Importantly, TCR stimulation
is universally required for cytokines to reprogramme
Tcell subsets35,65.
There are key differences in the requirements for
TCR and CD28 signalling in inflammatory and regu
latory Tcells. Although dispensable for the persis
tence of memory T helper cells, TReg cells (perhaps as
a consequence of self-antigen recognition) seem to be
ex-FOXP3 cells
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
Cytokine
receptor
Extracellular
cues
CTLA4
TCR
CD28
GLUT1
JAK
STAT
AKT
SOCS
Cytosolic
signalling and
metabolism
Glucose
PKM2
Glycolysis
Ca ,
MAPK
and IKK
OXPHOS
2+
Ribosome
Translation
PRC2
EZH2
Gene and
chromatin
regulation
STAT
K27me3
K27ac
HAT
Chromatin modiers,
cytokine receptors
and polarizing TFs
Polarizing cytokines,
chemokine receptors
and metabolic genes
NFAT
AP-1
NF-B
STAT
K9me3
ATP
Euchromatin:
accessible DNA
NFAT
AP-1
NF-B
Polarizing
cytokines
and TFs
HDAC
mRNA Nucleotides,
amino acids
Eector
and lipids
cytokines
K4me3
Polarizing TF
Heterochromatin:
inaccessible DNA
Figure 2 | The integration of signals at many levels in Tcells regulates plasticity. The regulation of Tcell
Nature
Reviews
| Immunology
plasticity is depicted at three levels in the cell: extracellular cues (red), cytosolic signalling and
metabolic
programmes
(orange) and transcription factor (TF)- or chromatin-mediated gene regulation (blue). These pathways are integrated by
mechanisms linking these levels of regulation. Direct proteinprotein interactions are indicated by solid lines, whereas
indirect links between proteins are denoted with dashed lines. AP1, activator protein 1; CTLA4, cytotoxic T lymphocyte
antigen 4; GLUT1, glucose transporter 1; HAT, histone acetyltransferase; HDAC, histone deacetylase; IKK, inhibitor of
nuclear factor-B kinase; JAK, Janus kinase; MAPK, mitogen-activated protein kinase; NFAT, nuclear factor of activated
Tcells; NF-B, nuclear factor-B; OXPHOS, oxidative phosphorylation; PKM2, pyruvate kinase M2; SOCS, suppressor of
cytokine signalling; STAT, signal transducer and activator of transcription; TCR, T cell receptor.
mTOR complex 1
(mTORC1). A complex
consisting of: mammalian
target of rapamycin (mTOR),
which is a serine/threonine
kinase; regulatory-associated
protein of mTOR (RAPTOR);
proline-rich AKT1 substrate of
40 kDa (PRAS40), which is an
mTORC1 inhibitor; mLST8,
which is of unknown function;
and DEP domain-containing
mTOR-interacting protein
(DEPTOR), which is an
mTORinhibitor.
REVIEWS
TH2
cell
IL-4
IL-2
TH9
cell
TH2/
TH17
cell
TH2/
TFH cell
pTReg
cell
TGF
TH2/
TH1 cell
Naive
CD4+ T cell
TR1
cell
TH17/
TFH cell
TH17
cell
IL-12,
IFN
TH1/
TFH cell
T H1
cell
TH1/
TH17
cell
IL-6
TFH
cell
IL-21
Polarization
Plasticity
mTORC2
A complex composed of
mammalian target of
rapamycin (mTOR), mLST8
andthe adaptor proteins
rapamycin-insensitive
companion of mTOR
(RICTOR)and stress-activated
MAP kinase-interacting
protein1 (SIN1).
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
Polarization index
TInam cell
CD28
IL-2R
TReg cell
TCR
GLUT1
PI3K
p85
p110
STAT5
LKB1
PIP2
PTEN
Glucose
PIP3
Ca2+
AMP/ATP
Glycolysis
PKM2
PDK
Nucleotides
AMPK
Amino acids
AKT
mTORC2
Pyruvate
Fatty acids
Lactate
ACC1
VHL
O2
HIF1
Acetyl-CoA
mTORC1
ATP
ETC
OXPHOS
TCA
cycle
Translation
FOXO
HIF1
Eector
cytokines
Glycolytic enzymes
Nucleus
Figure 4 | The PI3KAKTmTOR pathway and metabolic programmes converge to regulate the plasticity of
inflammatory versus regulatory Tcells. Links between extracellular cues, the phosphatidylinositol 3kinase (PI3K)
Nature Reviews
| Immunology
AKTmammalian target of rapamycin (mTOR) pathway, metabolic programmes and gene regulation
are depicted.
Apolarization index indicates proteins or processes that favour the induction of inflammatory (blue) or regulatory (red)
Tcell programmes. Direct proteinprotein interactions are indicated by solid lines, whereas indirect links between
proteins are denoted with dashed lines. ACC1, acetyl-CoA carboxylase 1; AMPK, AMP-activated protein kinase; ETC,
electron transport chain; FOXO, forkhead box O; GLUT1, glucose transporter 1; HIF1, hypoxia-inducible factor 1;
IL2R,IL2 receptor; LKB1, liver kinase B1; mTORC, mTOR complex; OXPHOS, oxidative phosphorylation; PDK,
phosphoinositide-dependent protein kinase; PIP2, phosphatidylinositol4,5biphosphate; PIP3, phosphatidylinositol3,4,5
trisphosphate; PKM2, pyruvate kinase M2; PTEN, phosphatase and tensin homologue; STAT5, signal transducer and
activator of transcription 5; TCA, tricarboxylic acid; TCR, T cell receptor; TInflam cell, inflammatory T cell; TReg cell, regulatory
Tcell; VHL, von Hippel-Lindau.
REVIEWS
Inaccessible
TF
GATA3
T-bet
Accessible
Polarization index
TReg cell
TInam cell
RORt
HDAC
BCL-6
PU.1
GATA3
FOXP3
RORt
BCL-6
GATA3
HP1
PRC1
K27ac
MBD
meCpG
K27me3
K9me3
PRC2
EZH2
CpG
DNMT
K9MT
G9A
SETDB1
SUV39H
KDM
lncRNA
HMT
BET
HAT
K4me3
me
TET
MLL
d
PTM
enzyme
Chromatin
remodeler
KDM
HMT
TF
TF
ub
ac
TF
p
me
Figure 5 | Mechanisms of gene regulation in Tcell plasticity. a | Direct interactions between transcription factors (TFs)
can antagonize the function of opposing TFs. b | DNA accessibility controls gene expression. Histone
modifications
and
Nature Reviews
| Immunology
DNA methylation can drive changes in chromatin structure, altering the accessibility of DNA to TFs. These epigenetic
changes act to stabilize gene expression programmes during Tcell responses in which driving or opposing TFs may be
transiently lost or gained, respectively. A polarization index indicates proteins or processes that favour inflammatory (blue)
or regulatory (red) Tcell programmes. c | TFs and long non-coding RNAs (lncRNAs) link chromatin modifiers to specific
genetic loci. d | Post-translational modifying (PTM) enzymes regulate the stability, activity and localization of TFs and may
provide a therapeutic means to indirectly target TF activity. ac, acetyl; BCL6, B cell lymphoma 6; BET, bromodomain and
extraterminal protein; DNMT, DNA methyltransferase; FOXP3, forkhead box P3; GATA3, GATA-binding protein 3; HAT,
histone acetyltransferase; HDAC, histone deacetylase; HMT, histone methyltransferase; HP1, heterochromatin protein 1;
KDM, histone lysine demethylase; K9MT, K9 methyltransferase; MBD, methyl-CpG-binding domain protein; me, methyl;
MLL, mixed-lineage leukaemia; p, phosphate; PRC, polycomb repressive complex; RORt,retinoic acid receptor-related
orphan receptor-t; TET, ten-eleven translocation; TInflam cell, inflammatory T cell; TReg cell, regulatory T cell; ub, ubiquitin.
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
cues to carry out the induction of a defined set of gene
effectors. Importantly, these transcription factors are
also expressed by innate lymphoid cells to engender
similar phenotypic traits118. Third, the expression of
specific STATs and master transcription factors is not
sufficient for polarization or plasticity of T helper cell
subsets; rather, a collection of additional transcription
factors are required119,120 (FIG.1). Additionaltranscrip
tion factors, such as the nuclear receptor 4A (NR4A)
family, transcription regulator protein BACH2, RUNX
proteins, retinoic acid receptors and aryl hydrocar
bon receptor (AHR), clearly have essential roles in the
maintenance of polarized states, as the disruption of
these transcription factors leads to enhanced plasticity
between subsets43,121125. Nevertheless, even the expres
sion of a suite of transcription factors is not sufficient
to drive the transcriptional programmes of T helper
cell subsets because access of the transcription factors
to their DNA-binding sites in the genome is regulated.
DNA accessibility by modification of DNA and histones.
In the nucleus, a delicate balance is struck between
the logistics of packing DNA into this confined space
(inheterochromatin) and providing access to DNA for
transcription (in euchromatin). Post-transcriptional
modifications of histone proteins, which make up nucleo
somes, control how tightly nucleosomes are packed, and
this has been adopted by the cell to regulate transcrip
tion in a semi-stable manner, along with the direct modi
fication of DNA by methylation. These forms of gene
regulation are termed epigenetic because they promote
the heritable transmission of distinct transcriptional
programmes despite identical DNA sequences.
REVIEWS
JMJD3 (also known as KDM6B), TH17 cell polarization
is enhanced, and many polarized Tcell subsets exhibit
enhanced stability 156. Finally, PRC1, which recognizes
H3K27me3 marks and further condenses chromatin,
is crucial for TH2 cell function and for limiting their
plasticity towards TH1 cell phenotypes136.
In contrast to H3K27 methylation, the regulation
of H3K9 methylation is more complex, owing to its
control by many distinct HMTs. This probably contrib
utes to the diversity of consequences observed with the
disruption of different H3K9 HMTs, as each enzyme
may interact with different binding partners or target
unique genomic loci. For example, deletion of H3K9
HMT SUV39H1 allows TH2polarized cells to express
IFN when restimulated in TH1 cellpolarizing condi
tions, whereas deficiency for HMT G9A (also known
as EHMT2) drives TReg cell and TH17 cell polarization
by opening the Foxp3 and Rorc loci157,158. In human
TRegcells, H3K9 HMT SETDB1 is coupled to FOXP3
by a FOXP3interacting KRAB domain-containing
protein (FIK)KRAB domain-associated protein (KAP;
also known as TIF1) complex to maintain repressive
chromatin states at the IL2 and IFNG loci159.
Global analyses of histone modifications associ
ated with transcriptionally accessible (H3K4me3) or
repressed (H3K27me3) loci in different polarized Tcell
subsets provide further mechanistic insight into how
Tcells can both acquire specific functions yet retain
plasticity. Whereas the cytokine loci of different Tcell
subsets exhibit either H3K4me3 or H3K27me3 marks,
the chromatin structure at most polarizing transcrip
tion factor loci contains both marks, indicative of a
permissive (or poised) chromatin state, allowing for
the induction of transcription factors from opposing
subsets to initiate cellular reprogramming 117. However,
as these studies must be performed on pooled cells, it
is important to consider that mixed or bivalent marks
at given loci may also represent heterogeneity in cell
populations or even allelic variability within cells11.
Importantly, the activities of chromatin-modifying
enzymes are responsive to changes in the environ
ment, exhibiting direct links to cytokine, metabolic
and kinase signalling pathways152,160162 (FIG.2). Overall,
it is clear that most chromatin- and DNA-modifying
enzymes have important roles in all CD4+ Tcell subsets,
acting to stabilize transcriptional programmes and cell
functions, often through cooperation with the polar
izing transcription factors or lncRNAs in each subset.
However, based on the current, albeit limited, state of
investigations so far, it seems that inflammatory and
regulatory programmes rely more heavily on different
chromatin modifiers, which leaves open the possibility
that fine-tuned inhibition of epigenetic enzymes might
selectively drive d
ifferent Tcell functions.
microRNAs. Although not covered here, it is important
to consider that Tcell plasticity is regulated by micro
RNA-mediated post-transcriptional regulation of a wide
variety of genes such as those involved in cytokinesig
nalling, TCR and costimulatory signalling, cytosolic
signalling and transcriptional regulatory pathways163.
Plasticity in disease
Plasticity is widely observed in immune-based dis
eases, such as autoimmunity and cancer. Patients with
various forms of autoimmune disease, such as type1
diabetes, multiple sclerosis or juvenile arthritis, exhibit
reduced stability of FOXP3 expression in TReg cells
and/or increased proportions of IFN-producing
FOXP3+ TRegcells14,48,164166. In individuals with food
allergies, TReg cells have characteristics of TH2 cells,
producing IL4 (REF.167). Plasticity towards TH17 cell
phenotypes is associated with several diseases, such
as IL17+FOXP3+TReg cells in rheumatoid arthritis168,
RORt+FOXP3+ TReg cells producing IL17 in colo
rectal cancer 169 and IL17expressing TH2 cells in atopic
asthma13. Therefore, it stands to reason that unique
microenvironments that are created in these diseases may
instigate Tcell reprogramming or that reprogrammed
Tcells may contribute to disease pathology. Indeed, in
each of the examples described above, reprogrammed
Tcells could contribute to mouse models of the human
diseases13,26,71,167169. In addition, in mouse models of
multiple sclerosis and inflammatory bowel disease, the
transition of TH17 cells to a TH1 or mixed TH17/TH1 cell
phenotype is necessary to drive the disease19,25,122,170,171.
In patients with human Tcell leukaemia virus type1
(HTLV1)associated myelopathy (also known as tropi
cal spastic paraparesis), a neuroinflammatory disorder
with multiple sclerosislike symptoms resulting from
infection with HTLV1, the virus is most likely to drive
disease by selectively infecting TReg cells and converting
them into IFN-producing TH1like cells by the direct
induction of Tbet expression by virally encoded pro
teins172. Thus, Tcell plasticity is an important factor in
immunological diseases, and the capacity to control Tcell
programming could lead to new therapies that ameliorate
disease by p
reventing deleterious, or promoting desirable,
Tcell functions.
Harnessing plasticity for immune therapy
Extracellular cues. Therapeutic targeting of the cues
that drive the polarization or function of CD4+ Tcells
is actively being pursued for the modulation of immu
nity. Targeting IL6, which is a crucial bifurcating signal
between TH17 and TReg cell polarization, with tocili
zumab, an antibody against the IL6 receptor, dampens
inflammation. Modulation of IL2induced signalling
can tip the balance between inflammatory Tcells and
TReg cells, such as has been demonstrated in humans
with low dose IL2 (REFS54,173). Alternatively, modu
lation of cytokine conformation, either by introduction
of synthetic mutant cytokines or by antibodies that bind
and alter cytokine conformation, may allow for selective
cytokine signalling to specific Tcell subsets, as recently
demonstrated for IL2 (REF.173). TCR signal strength
may also underlie the selective capacity of CD3specific
antibody treatment to deplete effector Tcells while
sparing TReg cells, delaying the progression of type1
diabetes174,175. Modulating CD28 co-stimulation with
antibodies blocking cytotoxic T lymphocyte antigen 4
(CTLA4) or with CTLA4Ig fusion constructs can also
drive immunity or tolerance, respectively 176.
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
Signalling cascades. The capacity to target key signal
ling cascades in Tcells that control plasticity is aided
by the development of many orally available kinase
inhibitors. Downstream of the cytokine receptors, small
molecule inhibitors of JAKs have proven to be success
ful as potent immunosuppressants, but their ability to
target specific T helper cell activities is limited because
several cytokine receptors use each kinase34. Blockade
of mTOR activity with rapamycin in type1 diabetes is
already under investigation in combination with IL2
to promote TReg cell function177. However, enhanced
TRegcell frequencies with rapamycin and IL2 is tran
sient because, as discussed, it seems that inhibition
further up the signalling cascade at AKT is most impor
tant for the promotion of TReg cell stability and func
tion79,80. Importantly, inhibition of specific isoforms
of PI3K may have differential effects on inflammatory
versus regulatory cells, as inhibition of the PI3K sub
unit p110 selectively disrupts TReg cell stability and
promotes anticancer immunity 178. In addition to direct
administration to patients, inhibitors can be used dur
ing invitro expansion of cells for adoptive cell transfer
therapies to improve the functional stability of Tcells
after transfer into patients179. With the advent of chimeric
antigen receptor (CAR) technologies to redirect cytotoxic
Tcells to specific antigenic targets comes the potential
to engineer CARs that are linked to different intracellu
lar signalling domains that promote distinct functions
in CD4+ Tcells180,181.
Enforcing specific metabolic programmes can
redirect Tcell responses to ameliorate autoimmune
disease, such as blocking fatty acid synthesis by inhib
iting ACC1 with soraphen A, which promotes TReg cell
over TH17 cell functions111. Blocking glycolysis with
non-metabolizable glucose or with the AMPK agonist
metformin promotes T follicular regulatory cell over
TFHcell polarization, reversing the severity of lupus182.
BET inhibitors
Inhibitors that bind the
bromodomain and
extraterminal (BET) motif
ofseveral bromodomaincontaining proteins (BRDs),
blocking their interaction with
acetylated lysines on histones
and preventing their promotion
of transcription.
REVIEWS
are revealing because they indicate that the retention
of phenotypic flexibility in Tcells is rooted in selective
advantages to thehost.
Concluding remarks
Tcells polarize in response to different pathogens in the
context of the unique microenvironments they create,
establishing stably directed Tcell responses, especially
at sites of infection such as tissues. However, Tcells
also retain the capacity to mould their function anew
upon reactivation in new polarizing environments.
Cytokines and transcription factors can drive repro
gramming, but they must do so within the confines of
the cytosolic and epigenetic circuitry that is established
in the cell to stabilize polarized Tcell functions during
effector responses. Several key regulatory nodes that
broadly divert Tcell functions towards inflammatory
or regulatory capacities are now apparent, suggesting
that modulating immunity by reprogramming Tcell
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
48. McClymont,S.A. etal. Plasticity of human regulatory
Tcells in healthy subjects and patients with type1
diabetes. J.Immunol. 186, 39183926 (2011).
49. Campbell,D.J. & Koch,M.A. Phenotypical and
functional specialization of FOXP3+ regulatory Tcells.
Nat. Rev. Immunol. 11, 119130 (2011).
50. Feng,T., Cao,A.T., Weaver,C.T., Elson,C.O. &
Cong,Y. Interleukin12 converts Foxp3+ regulatory
Tcells to interferon--producing Foxp3+ Tcells that
inhibit colitis. Gastroenterology 140, 20312043
(2011).
51. Fuhrman,C.A. etal. Divergent phenotypes of human
regulatory T cells expressing the receptors TIGIT and
CD226. J.Immunol. 195, 145155 (2015).
52. Tang,Q. etal. Central role of defective interleukin2
production in the triggering of islet autoimmune
destruction. Immunity 28, 687697 (2008).
53. Feng,Y. etal. Control of the inheritance of regulatory
Tcell identity by a cis element in the Foxp3 locus.
Cell158, 749763 (2014).
54. Klatzmann,D. & Abbas,A.K. The promise of lowdoseinterleukin2 therapy for autoimmune and
inflammatory diseases. Nat. Rev. Immunol. 15,
283294 (2015).
55. van Panhuys,N., Klauschen,F. & Germain,R.N.
Tcell-receptor-dependent signal intensity dominantly
controls CD4+ T cell polarization invivo. Immunity 41,
6374 (2014).
56. Yamane,H. & Paul,W.E. Early signaling events that
underlie fate decisions of naive CD4+ Tcells toward
distinct Thelper cell subsets. Immunol. Rev. 252,
1223 (2013).
57. Fazilleau,N., McHeyzer-Williams,L.J., Rosen,H. &
McHeyzer-Williams,M.G. The function of follicular
helper Tcells is regulated by the strength of Tcell
antigen receptor binding. Nat. Immunol. 10,
375384 (2009).
58. Tubo,N.J. etal. Single naive CD4+ T cells from a
diverse repertoire produce different effector cell
typesduring infection. Cell 153, 785796 (2013).
59. Sauer,S. etal. Tcell receptor signaling controls Foxp3
expression via PI3K, Akt, and mTOR. Proc. Natl Acad.
Sci. USA 105, 77977802 (2008).
60. Gottschalk,R.A., Corse,E. & Allison,J.P. TCR ligand
density and affinity determine peripheral induction of
Foxp3 invivo. J.Exp. Med. 207, 17011711 (2010).
61. Hsieh,C.S., Lee,H.M. & Lio,C.W. Selection of
regulatory Tcells in the thymus. Nat. Rev. Immunol.
12, 157167 (2012).
62. Ohkura,N. etal. Tcell receptor stimulation-induced
epigenetic changes and Foxp3 expression are
independent and complementary events required
forTreg cell development. Immunity 37, 785799
(2012).
This study identifies key epigenetic features of
TRegcells driven by TCR signals and independent
ofFOXP3 expression.
63. Nagaoka,M., Hatta,Y., Kawaoka,Y. & Malherbe,L.P.
Antigen signal strength during priming determines
effector CD4 T cell function and antigen sensitivity
during influenza virus challenge. J.Immunol. 193,
28122820 (2014).
64. Ahmadzadeh,M. & Farber,D.L. Functional plasticity
of an antigen-specific memory CD4 Tcell population.
Proc. Natl Acad. Sci. USA 99, 1180211807 (2002).
65. Martinez-Sanchez,M.E., Mendoza,L., Villarreal,C.
&Alvarez-Buylla,E.R.A. Minimal regulatory network
of extrinsic and intrinsic factors recovers observed
patterns of CD4+ T cell differentiation and plasticity.
PLoS Comput. Biol. 11, e1004324 (2015).
66. Moran,A.E. etal. Tcell receptor signal strength in
Treg and iNKT cell development demonstrated by a
novel fluorescent reporter mouse. J.Exp. Med. 208,
12791289 (2011).
67. Levine,A.G., Arvey,A., Jin,W. & Rudensky,A.Y.
Continuous requirement for the TCR in regulatory
Tcell function. Nat. Immunol. 15, 10701078
(2014).
68. Vahl,J.C. etal. Continuous T cell receptor signals
maintain a functional regulatory T cell pool. Immunity
41, 722736 (2014).
69. Tang,Q. etal. Cutting edge: CD28 controls peripheral
homeostasis of CD4+ CD25+ regulatory Tcells.
J.Immunol. 171, 3348 (2003).
70. Zhang,R. etal. An obligate cell-intrinsic function
forCD28 in Tregs. J. Clin. Invest. 123, 580593
(2013).
71. Bailey-Bucktrout,S.L. etal. Self-antigen-driven
activation induces instability of regulatory T cells
during an inflammatory autoimmune response.
Immunity 39, 949962 (2013).
REVIEWS
121. Roychoudhuri,R. etal. BACH2 represses effector
programs to stabilize Treg-mediated immune
homeostasis. Nature 498, 506510 (2013).
122. Wang,Y. etal. The transcription factors Tbet and
Runx are required for the ontogeny of pathogenic
interferon-producing T helper 17 cells. Immunity 40,
355366 (2014).
123. Sekiya,T. etal. Suppression of Th2 and Tfh immune
reactions by Nr4a receptors in mature T reg cells.
J.Exp. Med. 212, 16231640 (2015).
This paper reveals the essential role of additional
transcription factors that collaborate with FOXP3
to maintain the TReg cell transcriptional programme.
124. Brown,C.C. etal. Retinoic acid is essential for Th1 cell
lineage stability and prevents transition to a Th17 cell
program. Immunity 42, 499511 (2015).
125. Park,B.V. & Pan,F. The role of nuclear receptors in
regulation of Th17/Treg biology and its implications for
diseases. Cell. Mol. Immunol. 12, 533542 (2015).
126. Bird,J. etal. Helper Tcell differentiation is controlled
by the cell cycle. Immunity 9, 229237 (1998).
127. Agarwal,S. & Rao,A. Modulation of chromatin
structure regulates cytokine gene expression during
Tcell differentiation. Immunity 9, 765775 (1998).
128. Grogan,J.L. etal. Early transcription and silencing of
cytokine genes underlie polarization of T helper cell
subsets. Immunity 14, 205215 (2001).
129. Avni,O. etal. TH cell differentiation is accompanied
bydynamic changes in histone acetylation of cytokine
genes. Nat. Immunol. 3, 643651 (2002).
130. Hojfeldt,J.W., Agger,K. & Helin,K. Histone lysine
demethylases as targets for anticancer therapy.
Nat.Rev. Drug Discov. 12, 917930 (2013).
131. Pastor,W.A., Aravind,L. & Rao,A. TETonic shift:
biological roles of TET proteins in DNA demethylation
and transcription. Nat. Rev. Mol. Cell Biol. 14,
341356 (2013).
132. Zhang,F. T helper type1specific Brg1 recruitment
and remodeling of nucleosomes positioned at the
IFNpromoter are Stat4 dependent. J.Exp. Med.
203, 14931505 (2006).
133. Schones,D.E. etal. Dynamic regulation of
nucleosome positioning in the human genome.
Cell132, 887898 (2008).
134. Wang,Z. etal. Genome-wide mapping of HATs and
HDACs reveals distinct functions in active and inactive
genes. Cell 138, 10191031 (2009).
135. Ichiyama,K. etal. The methylcytosine dioxygenase
Tet2 promotes DNA demethylation and activation
ofcytokine gene expression in T Cells. Immunity 42,
613626 (2015).
136. Onodera,A. & Nakayama,T. Epigenetics of Tcells
regulated by Polycomb/Trithorax molecules.
TrendsMol. Med. 21, 330340 (2015).
137. Miller,S.A., Huang,A.C., Miazgowicz,M.M.,
Brassil,M.M. & Weinmann,A.S. Coordinated but
physically separable interaction with H3K27
demethylase and H3K4methyltransferase activities
are required for Tbox protein-mediated activation
ofdevelopmental gene expression. GenesDev. 22,
29802993 (2008).
138. Bettini,M.L. etal. Loss of epigenetic modification
driven by the Foxp3 transcription factor leads to
regulatory T cell insufficiency. Immunity 36, 717730
(2012).
139. Wang,L. etal. Mbd2 promotes Foxp3 demethylation
and Tregulatory-cell function. Mol. Cell. Biol. 33,
41064115 (2013).
140. Yang,R. etal. Hydrogen sulfide promotes Tet1andTet2mediated Foxp3 demethylation to drive
regulatory T cell differentiation and maintain immune
homeostasis. Immunity 43, 251263 (2015).
References 139 and 140 reveal that DNA
demethylation by TET proteins is crucial for the
maintenance of FOXP3 expression and TReg cell
stability.
141. Liu,Y. etal. Two histone/protein acetyltransferases,
CBP and p300, are indispensable for Foxp3+
Tregulatory cell development and function.
Mol.Cell.Biol. 34, 39934007 (2014).
142. Yamashita,M. etal. Crucial role of MLL for the
maintenance of memory T helper type2 cell
responses. Immunity 24, 611622 (2006).
143. Collier,S.P., Collins,P.L., Williams,C.L.,
Boothby,M.R. & Aune,T.M. Cutting edge: influence
of Tmevpg1, a long intergenic noncoding RNA, on
theexpression of Ifng by Th1 cells. J.Immunol. 189,
20842088 (2012).
144. Gomez,J.A. etal. The NeST long ncRNA controls
microbial susceptibility and epigenetic activation of
the interferon- locus. Cell 152, 743754 (2013).
www.nature.com/nri
2016 Macmillan Publishers Limited. All rights reserved
REVIEWS
192. Fu,W. etal. Epigenetic modulation of type1
diabetesvia a dual effect on pancreatic macrophages
and cells. eLife 3, e04631 (2014).
References 190192 collectively show that
targeting epigenetic enzymes with systemic drugs
can have selective immunomodulatory activities
toameliorate cancer or autoimmune disease.
193. Tsuji,M. etal. Preferential generation of follicular
Bhelper Tcells from Foxp3+ Tcells in gut Peyers
patches. Science 323, 14881492 (2009).
194. Sharma,M.D. etal. An inherently bifunctional
subsetof Foxp3+ T helper cells is controlled by the
transcription factor Eos. Immunity 38, 9981012
(2013).
195. Neumann,C. etal. Role of Blimp1 in programing
Theffector cells into IL10 producers. J.Exp. Med.
211, 18071819 (2014).
196. Esplugues,E. etal. Control of TH17 cells occurs in
thesmall intestine. Nature 475, 514518 (2011).
Acknowledgements